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In Vitro Expression of Pigeonpea Lectin Gene and its Characterization

KrishiKosh

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Title In Vitro Expression of Pigeonpea Lectin Gene and its Characterization
 
Creator SUDHIR KUMAR
 
Contributor Rekha Kansal
 
Subject null
 
Description t-8994
Crop losses due to insect pests are a significant factor in limiting food
production. Worldwide use of chemical insecticides and pesticides has increased the cost
of pest control, resulted in insecticidal hazards to biological organisms, pollution to the
environment and increasing insect resistance. Therefore, the challenge today is to
achieve higher and stable crop production with safe and eco-friendly strategies. Plants
accumulate a set of defense proteins including lectins, proteinase inhibitors, amylase
inhibitors etc. Lectins reversibly and non-enzymatically bind specific carbohydrates and
this agglutination property makes them useful against various lepidopteran and
homopteran insect pests. The harmful effects of lectins on biological parameters of
insects include loss in weight, mortality, feeding inhibition, delays in total
developmental duration, adult emergence and fecundity on the first and second
generation. Realizing their importance, lectin gene have been isolated and characterized
from pigeon pea. The isolated pigeonpea lectin (PPL) gene (~825 bp) was first cloned in
pENTR-D-TOPO vector, subcloned into an expression vector (Gateway Destination
vector pET300/NT-DEST) and transformed into BL21 DE3 pLysS competent cells of E.
coli for protein expression studies. The PPL gene expression studies were carried out at
different temperatures, IPTG concentrations and time intervals. The expression was
maximum at 2.0 and 2.50 mM IPTG concentration at 37 ºC for 5 hrs. The size of the
protein was found to be around ~30 KDa, as predicted by using the ProtParam software.
The expression was confirmed by SDS-PAGE and western blotting. Thus, transferring
these defense genes under the control of tissue specific promoters will be an effective
tool for sustainable insect pest management programme.
 
Date 2017-06-08T05:25:05Z
2017-06-08T05:25:05Z
2014
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/5810018706
 
Language en_US
 
Format application/pdf
 
Publisher Division of Molecular Biology and Biotechnology Indian Agricultural Research Institute New Delhi